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<button class="collapsible">Electrochemistry 3-Electrode Setup</button> | <button class="collapsible">Electrochemistry 3-Electrode Setup</button> | ||
<div class="drop"> | <div class="drop"> | ||
− | < | + | <h5>Motivation</h5> |
+ | |||
+ | <p>Current electrochemistry setup can cost over £1000 as they include a counter and working electrodes made of materials such as gold as well as AgCl reference electrodes. While this kind of set-up has its own advantages such as great biocompatibility, outstanding conductivity and low reactivity with organic molecules, pure gold wires are too expensive to make it affordable and cost-effective. Instead, we opted for carbon electrodes which are up to 7 times cheaper than gold wire when comparing price per cm<sup>2</sup> - the main geometric feature of interest in electrochemistry set-ups and 250 times cheaper when looking at the three electrodes required to stimulate gene expression. <p> | ||
+ | <div class="center"> | ||
+ | https://static.igem.org/mediawiki/2018/c/c3/T--Imperial_College--electrodesetup.jpg | ||
+ | </div> | ||
+ | |||
+ | <div class="center"> | ||
+ | https://static.igem.org/mediawiki/2018/c/c3/T--Imperial_College--electrodesetup.jpg | ||
+ | </div> | ||
+ | |||
+ | <h5>Characteristics & Performance </h5> | ||
+ | <p>To characterise our system a series of square wave voltammetry and cyclic voltammetry experiments were performed. For these experiments, a 90% content carbon working electrode of 2mm diameter was used, a 60% carbon counter electrode of 5mm diameter and a custom-made Ag/AgCl reference electrode was used (protocolHYPERLINK). | ||
+ | |||
+ | To drive the specific redox potential into our system amperometry experiments were performed. For these, the setup only differs in the use of 60% carbon working electrodes of 5mm diameter as opposed to 90% content carbon working electrode of 2mm diameter. | ||
+ | |||
+ | A table for the technical specifications of these electrodes is found here:<p> | ||
+ | |||
+ | <table class="tg"> | ||
+ | <tr> | ||
+ | <td class="tg-cnwr">Electrode number</td> | ||
+ | <td class="tg-cnwr">Electrode type</td> | ||
+ | <td class="tg-cnwr">Length (mm)</td> | ||
+ | <td class="tg-cnwr">Diameter(mm)</td> | ||
+ | <td class="tg-cnwr">Specifications</td> | ||
+ | <td class="tg-cnwr">Capacitance (S/m)</td> | ||
+ | </tr> | ||
+ | <tr> | ||
+ | <td class="tg-mfhl">1</td> | ||
+ | <td class="tg-mfhl">Working/Counter</td> | ||
+ | <td class="tg-mfhl">100</td> | ||
+ | <td class="tg-mfhl">5</td> | ||
+ | <td class="tg-mfhl">60% Carbon - 40% Epoxy Resin</td> | ||
+ | <td class="tg-mfhl">195 ± 11</td> | ||
+ | </tr> | ||
+ | <tr> | ||
+ | <td class="tg-mfhl">2</td> | ||
+ | <td class="tg-mfhl">Working</td> | ||
+ | <td class="tg-mfhl">100</td> | ||
+ | <td class="tg-mfhl">2</td> | ||
+ | <td class="tg-mfhl">90% Carbon - 5% Clay - 5% Wax</td> | ||
+ | <td class="tg-mfhl">796 ± 38</td> | ||
+ | </tr> | ||
+ | <tr> | ||
+ | <td class="tg-mfhl">3</td> | ||
+ | <td class="tg-mfhl">Reference</td> | ||
+ | <td class="tg-mfhl">10</td> | ||
+ | <td class="tg-mfhl">0.5</td> | ||
+ | <td class="tg-mfhl">99.9% Silver</td> | ||
+ | <td class="tg-mfhl">Ø</td> | ||
+ | </tr> | ||
+ | </table> | ||
+ | |||
+ | <p>This cheap 3-electrode setup was capable enough of stimulating gene expression as shown in figure XX -(THOMAS PICTURE OF GFP DOT) and figure XX (FIGURE OF VOLTAMMETRY) (FIGURE OF PAPER VOLTAMMETRY)<p> | ||
+ | |||
+ | <div class="center"> | ||
+ | https://static.igem.org/mediawiki/2018/c/c3/T--Imperial_College--electrodesetup.jpg | ||
+ | </div> | ||
+ | |||
+ | <div class="center"> | ||
+ | https://static.igem.org/mediawiki/2018/c/c3/T--Imperial_College--electrodesetup.jpg | ||
+ | </div> | ||
+ | |||
+ | <div class="center"> | ||
+ | https://static.igem.org/mediawiki/2018/c/c3/T--Imperial_College--electrodesetup.jpg | ||
+ | </div> | ||
+ | |||
+ | <h5>Price comparison</h5> | ||
+ | <h3>Comparison of price per setup</h3> | ||
+ | <p>TABLE HERE</p> | ||
+ | <h3>Comparison of price per cm<sup>2</sup></h3> | ||
+ | <p>TABLE HERE</p> | ||
+ | |||
</div> | </div> | ||
− | <button class="collapsible"> | + | <button class="collapsible">Programmable Electrode Array</button> |
<div class="drop"> | <div class="drop"> | ||
+ | |||
+ | <h5>Motivation</h5> | ||
+ | <p>Once we experimentally demonstrated how of gene expression could be locally induced with our affordable 3-electrode setup, the next goal was set to build a higher-end electrode array able to induce genetic expression locally at different regions in a lawn of cells. </p> | ||
+ | |||
+ | <h5>Concept Design</h5> | ||
+ | <p>The electrode array is printed onto one side of a Printed Circuit Board (PCB) in the form of 100 (10x10) circular working electrode pads, 4 counter rectangular pads, and 1 reference pad with the shape of a cross as it can be seen in figure XX ~(figure of pad configuration) . All the electronic components that allow to control the logic of the array are placed on the other side of the PCB. | ||
+ | We want each individual pad to select one of two working electrode potentials.The circuit can be broken down into several modules: | ||
+ | electrode cell : This corresponds to a memory storage unit, a set of two transistors and the electrode pad itself connected together. The storage unit is operated by a Data Flip-Flop and stores 1-bit (0 or 1),based on the data flip-flop output the transistors select one of the available Working potentials and outputs it into the pad. | ||
+ | Power module: | ||
+ | logic chips are powered through a voltage difference between a constant voltage supply (Vcc) and a ground(GND) supply. The logic HIGH (1) of a components corresponds to a voltage of Vcc while the logic LOW (0) corresponds to a voltage equal to GND. We want logic LOW to correspond to 2.5V below the reference voltage of the electrochemical array, and logic HIGH to correspond to 2.5V above the reference voltage. this is to ensure that the Data Flip-Flops can force the FETs into either an ON or OFF state. Thus, we cannot use real ground as logic GND. Instead, we run the logic (arduino and PCB chips) from a non-grounded battery, in order to be able to shift logic levels up or down. This shifting is achieved by an op-amp, with its non-inverting input connected to the reference electrode. The op-amp is connected in negative feedback to a 2.5V voltage regulator. This means that it sources and sinks current as required to maintain its output (2.5V relative to logic GND) at the reference voltage, effectively centering the logic HIGH and LOW around the reference voltage. This ensures that the logic circuits can reliably switch the FETs. | ||
+ | ROW-Selecting module: in order to program our electrode pads we need to sort through every single row and feed individual data into each pad. A demultiplexer selects electrode rows one at a time, telling each row in turn to read the data supplied to it. | ||
+ | Data-feeder: The data, which is either a 1 or a 0 (HIGH or LOW) is fed into the individual cells of the selected row. This operation is accomplished by a serial-input-parallel-output (SIPO) shift register which outputs 10 bits at a time (one per column of the electrode array).</p> | ||
<table class="tg"> | <table class="tg"> |
Revision as of 02:44, 18 October 2018
Hardware
Why do we need Hardware?
Hardware Requirments
PCB
Motivation
Current electrochemistry setup can cost over £1000 as they include a counter and working electrodes made of materials such as gold as well as AgCl reference electrodes. While this kind of set-up has its own advantages such as great biocompatibility, outstanding conductivity and low reactivity with organic molecules, pure gold wires are too expensive to make it affordable and cost-effective. Instead, we opted for carbon electrodes which are up to 7 times cheaper than gold wire when comparing price per cm2 - the main geometric feature of interest in electrochemistry set-ups and 250 times cheaper when looking at the three electrodes required to stimulate gene expression.
Characteristics & Performance
To characterise our system a series of square wave voltammetry and cyclic voltammetry experiments were performed. For these experiments, a 90% content carbon working electrode of 2mm diameter was used, a 60% carbon counter electrode of 5mm diameter and a custom-made Ag/AgCl reference electrode was used (protocolHYPERLINK). To drive the specific redox potential into our system amperometry experiments were performed. For these, the setup only differs in the use of 60% carbon working electrodes of 5mm diameter as opposed to 90% content carbon working electrode of 2mm diameter. A table for the technical specifications of these electrodes is found here:
Electrode number | Electrode type | Length (mm) | Diameter(mm) | Specifications | Capacitance (S/m) |
1 | Working/Counter | 100 | 5 | 60% Carbon - 40% Epoxy Resin | 195 ± 11 |
2 | Working | 100 | 2 | 90% Carbon - 5% Clay - 5% Wax | 796 ± 38 |
3 | Reference | 10 | 0.5 | 99.9% Silver | Ø |
This cheap 3-electrode setup was capable enough of stimulating gene expression as shown in figure XX -(THOMAS PICTURE OF GFP DOT) and figure XX (FIGURE OF VOLTAMMETRY) (FIGURE OF PAPER VOLTAMMETRY)
Price comparison
Comparison of price per setup
TABLE HERE
Comparison of price per cm2
TABLE HERE
Motivation
Once we experimentally demonstrated how of gene expression could be locally induced with our affordable 3-electrode setup, the next goal was set to build a higher-end electrode array able to induce genetic expression locally at different regions in a lawn of cells.
Concept Design
The electrode array is printed onto one side of a Printed Circuit Board (PCB) in the form of 100 (10x10) circular working electrode pads, 4 counter rectangular pads, and 1 reference pad with the shape of a cross as it can be seen in figure XX ~(figure of pad configuration) . All the electronic components that allow to control the logic of the array are placed on the other side of the PCB. We want each individual pad to select one of two working electrode potentials.The circuit can be broken down into several modules: electrode cell : This corresponds to a memory storage unit, a set of two transistors and the electrode pad itself connected together. The storage unit is operated by a Data Flip-Flop and stores 1-bit (0 or 1),based on the data flip-flop output the transistors select one of the available Working potentials and outputs it into the pad. Power module: logic chips are powered through a voltage difference between a constant voltage supply (Vcc) and a ground(GND) supply. The logic HIGH (1) of a components corresponds to a voltage of Vcc while the logic LOW (0) corresponds to a voltage equal to GND. We want logic LOW to correspond to 2.5V below the reference voltage of the electrochemical array, and logic HIGH to correspond to 2.5V above the reference voltage. this is to ensure that the Data Flip-Flops can force the FETs into either an ON or OFF state. Thus, we cannot use real ground as logic GND. Instead, we run the logic (arduino and PCB chips) from a non-grounded battery, in order to be able to shift logic levels up or down. This shifting is achieved by an op-amp, with its non-inverting input connected to the reference electrode. The op-amp is connected in negative feedback to a 2.5V voltage regulator. This means that it sources and sinks current as required to maintain its output (2.5V relative to logic GND) at the reference voltage, effectively centering the logic HIGH and LOW around the reference voltage. This ensures that the logic circuits can reliably switch the FETs. ROW-Selecting module: in order to program our electrode pads we need to sort through every single row and feed individual data into each pad. A demultiplexer selects electrode rows one at a time, telling each row in turn to read the data supplied to it. Data-feeder: The data, which is either a 1 or a 0 (HIGH or LOW) is fed into the individual cells of the selected row. This operation is accomplished by a serial-input-parallel-output (SIPO) shift register which outputs 10 bits at a time (one per column of the electrode array).
Part number | Part description | Required units |
GBP/unit | Total price (GBP) | Used Supplier | Manufacturer | Manufacturer Part number |
---|---|---|---|---|---|---|---|
1 | Printed Circuit Board (Gold-plated) | 1 | 67.69 | 67.79 | Beta Layout Inc | preferred | none |
2 | 4-to-16 Demultiplexer | 1 | 1.33 | 1.33 | DigiKey | Nexperia USA Inc. | 74HCT4514D,653 |
3 | 12 Output Serial-Input-Parallel-Output Shift Register |
1 | 1.32 | 1.32 | DigiKey | Nexperia USA Inc. | NPIC6C4894PW-Q100J |
4 | Single Output Data Flip Flop | 100 | 0.103 | 10.3 | Farnell Element 14 | NEXPERIA | 74AHC1G79GW,125 |
5 | Dual N-P MOSFET | 100 | 0.265 | 26.5 | Farnell Element 14 | NEXPERIA | PMCPB5530X |
6 | Operational Amplifier | 1 | 0.967 | 0.97 | Farnell Element 14 | Texas Instruments | LMV651MG |
7 | 2.5 Volts - Voltage Regulator | 1 | 1.2 | 1.2 | DigiKey | Texas Instruments | LP38690SD-2.5/NOPB |
8 | 1 \μF Capacitor | 2 | 0.058 | 0.12 | Farnell Element 14 | TDK | C1608X5R1C105K080AA |
9 | 100 nF Capacitor | 100 | 0.103 | 10.3 | Farnell Element 14 | MURATA | GCJ188R71H104KA12D |
10 | 0 \Ω Resistor | 1 | 0.0273 | 0.27 | Farnell Element 14 | Panasonic Electronic Components |
ERJ3GEY0R00V |
Total | Programmable 10x10 Electrode Array |
1 | / | 120.1 | PixCell Electronics Team |
/ | / |
Part number | Part description | Link |
---|---|---|
1 | Printed Circuit Board (Gold-plated) | Beta Layout |
2 | 4-to-16 Demultiplexer | DigiKey - Shift Register |
3 | 12 Output Serial-Input-Parallel-Output Shift Register |
DigiKey - Demultiplexer |
4 | Single Output Data Flip Flop | Farnell - FlipFlop |
5 | Dual N-P MOSFET | Farnell - MOSFET |
6 | Operational Amplifier | Farnell - Op-Amp |
7 | 2.5 Volts - Voltage Regulator | DigiKey - 2.5VR |
8 | 1 \uF Capacitor | Farnell - C_1\u |
9 | 100 nF Capacitor | Farnell - 100nF |
10 | 0 \Ohm Resistor | Farnell - R_0\Ohm |
Total | Programmable 10x10 Electrode Array |